Single-Stranded Antisense siRNAs Guide Target RNA Cleavage in RNAi

  title={Single-Stranded Antisense siRNAs Guide Target RNA Cleavage in RNAi},
  author={Javier Ruiz Martinez and Agnieszka Patkaniowska and Henning Urlaub and Reinhard L{\"u}hrmann and Thomas Tuschl},
Small interfering RNAs (siRNAs) are the mediators of mRNA degradation in the process of RNA interference (RNAi). Here, we describe a human biochemical system that recapitulates siRNA-mediated target RNA degradation. By using affinity-tagged siRNAs, we demonstrate that a single-stranded siRNA resides in the RNA-induced silencing complex (RISC) together with eIF2C1 and/or eIF2C2 (human GERp95) Argonaute proteins. RISC is rapidly formed in HeLa cell cytoplasmic extract supplemented with 21 nt… CONTINUE READING
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